*Target (protein/gene name): AcpA (secreted acid phosphatase) *NCBI Gene # or RefSeq#: FTN_0090 *Protein ID (NP or XP #) or Wolbachia#: YP_897755.1 *Organism (including strain): Francisella tularensis Etiologic Risk Group: Those who live or visit areas of Arkansas, Missouri, or Oklahoma due to the concentration of ticks. Also those who engage in hunting, trapping, gardening, landscaping, or work in wildlife management or veterinary medicine. *Disease Information (sort of like the Intro to your Mini Research Write up): Tularemia is an infectious disease caused by the bacteria Francisella tularensis, commonly found in animals such as rodents, rabbits and hares [4]. Humans can contract tularemia by means of getting bitten by an infected tick, deerfly, or other insect, handling an infected animal carcass, eating or drinking contaminated food or water, or breathing in the bacteria itself [4]. Tularemia ranges from mild to life-threatening, often with the accompanying symptoms of a sudden fever, chills, headaches, diarrhea, muscle aches, joint paint, dry cough, and progressive weakness [4]. More serious symptoms include development of pneumonia, chest pain, cough and difficulty breathing [4]. Tularemia occurs naturally in the United States and because of its highly infectious nature, it can be used as a bioterrorism weapon. Link to TDR Targets page (if present): n/a Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) http://www.ncbi.nlm.nih.gov/gene/?term=FTN_0090 Essentiality of this protein: Essential in producing, acquiring, and mobilizing inorganic phosphates in the cell signaling transduction pathway Is it a monomer or multimer as biological unit? (make prediction athttp://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): 2-mer Complex of proteins?: complexed with othrovanadate (2D1G) Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): No evidence showing specific druggability of AcpA in another organism, but there are papers showing druggability of secreted phosphatases in other organisms
*EC#: 3.1.3.2 Link to BRENDA EC# page: http://www.brenda-enzymes.org/enzyme.php?ecno=3.1.3.2&Suchword=&organism%5B%5D=Francisella+tularensis -- Show screenshot of BRENDA enzyme mechanism schematic Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
A discontinuous colorimetric assay was used. Standard 0.2 ml assay mixture contained 0.2 M sodium acetate, pH 6.0, 2.0 mM p-nitrophenylphosphate (pNPP), and varying amounts of enzyme. Mixtures incubated at 37oC for 15 minutes, and reaction was stopped by addition of 0.2 ml of 0.5 M glycine, pH 10. -- link to Sigma (or other company) page for assay (see Sigma links below) -- -or link (or citation) to paper that contains assay information http://www.sciencedirect.com/science/article/pii/S1046592805001580 -- links to assay reagents (substrates) pages. http://www.sigmaaldrich.com/catalog/product/sigma/g7126?lang=en®ion=US --- List cost and quantity of substrate reagents, supplier, and catalog # Glycine
Several different sizes available, see site above for complete list.
Supplier:ReagentPlus®
10 mg = $17.80
100 g = $23.50
1 kg = $64.50
Structure (PDB or Homology model) -- PDB # or closest PDB entry if using homology model: 2D1G
Current Inhibitors:
AgNO3, Arsenate, Cu2+, FeSO4, Hg2+, Molybdate, Orthovanadate, Tartrate, Tungstate, Vanadate, Zn2+ Expression Information (has it been expressed in bacterial cells): Expressed in E. coli BL21 (DE3) Purification Method: SP-Sepharose cation exchange resin (25 ml) Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): (NOT INCLUDING ADDED X)
MKLNKITLGILSLSIATTTFATDVNNSKPNDYGTLVKIEQKLFNNANTLKTTTPIKHVVIIFQENNSFDRYFGMYPN
AKNPEGEPKFVAKENTPNVNGLTKQLLENNPNTKNPYRLDRNFQPCSQNHEYHQEISSFNGGLMNKFVEHG
GHDNDTYKQNCDGQVMGYYDGNTVTALWNYAQNFALNDNTFGTTFGPSTPGALNLVAGANGPAMSPSGNL
ENIENNYIIDDPNPYYDDCSYGTSKSGDTNTAVAKITDGYNIGHYLTQKGITWGWFQGGFKPTSYSGKTAICDA
MSTNKFGVKSRDYIPHHEPFNYWKETSNPHHLAPSDDKYIGSNDQANHQYDISEFWKALDQNNMPAVSYLKA
PGYQDGHGGYSNPLDEQEWLVNTINRIQQSKDWDSTAIIIIYDDSDGDYDHVYSPKSQFSDIKGRQGYGPRLP
MLVISPYAKANYVDHSLLNQASVLKFIEYNWGIGSVSKYSNDKYSNNILNMFDFNKEQKTLKLILDPKTGLVMD
KLN *length of your protein in Amino Acids: 514
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website:
57.78 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 91930
Abs 0.1% (=1 g/l) 1.591, assuming all pairs of Cys residues form cystines
Ext. coefficient 91680
Abs 0.1% (=1 g/l) 1.587, assuming all Cys residues are reduced
This present two possible trans-membrane regions and one definite region. Therefore there is 1-3 trans-membrane regions.
*CDS Gene Sequence (paste as text only): ATGGACGTGAATAATTCTAAGCCGAACGATTACGGTACCCTGGTTAAAATCGAACAGAAACTGTTTAACAACGCGAATACGCTCAAAACGACCACCCCGATTAAACACGTAGTCATTATCTTCCAGGAGAATAACTCTTTTGATCGTTACTTCGGTATGTACCCGAATGCGAAGAATCCGGAAGGTGAACCAAAGTTTGTTGCGAAAGAAAACACCCCAAATGTTAACGGCCTCACCAAACAGCTGCTCGAAAATAACCCGAACACCAAAAATCCGTACCGCCTGGATCGCAACTTCCAGCCGTGCTCTCAGAACCACGAATACCACCAGGAAATCTCTTCTTTCAATGGTGGCCTGATGAATAAGTTCGTCGAACATGGTGGTCACGACAATGACACCTACAAACAGAATTGCGATGGTCAGGTGATGGGCTACTATGATGGCAATACTGTTACCGCGCTGTGGAATTACGCGCAGAACTTCGCGCTGAATGACAACACCTTTGGTACTACCTTCGGTCCGTCTACCCCGGGTGCCCTGAACCTGGTTGCGGGTGCAAACGGTCCGGCGATGTCTCCGTCTGGCAACCTCGAGAACATCGAAAACAACTACATCATTGACGACCCTAACCCGTATTATGACGACTGCTCTTACGGCACCTCTAAATCTGGCGACACCAACACTGCGGTTGCTAAGATCACGGATGGTTACAACATCGGTCACTACCTCACCCAAAAAGGTATCACTTGGGGTTGGTTTCAAGGCGGCTTCAAACCGACCTCTTACTCTGGTAAGACGGCGATTTGTGACGCCATGTCTACCAACAAGTTCGGTGTTAAATCTCGTGACTATATCCCGCACCACGAGCCTTTCAACTATTGGAAGGAGACCTCCAATCCACACCACCTCGCGCCGAGCGACGACAAATACATTGGTTCTAATGATCAAGCGAATCACCAATACGACATCTCTGAATTCTGGAAGGCGCTCGACCAGAACAACATGCCTGCCGTTTCTTACCTGAAAGCTCCGGGTTACCAGGATGGCCACGGTGGCTACTCCAACCCACTGGACGAACAGGAATGGCTCGTTAATACCATCAACCGTATCCAGCAGTCTAAAGACTGGGACTCTACCGCTATTATCATCATTTACGATGACAGCGACGGTGATTACGATCACGTTTACTCTCCAAAGTCTCAATTCTCTGACATCAAGGGCCGTCAAGGTTATGGTCCGCGTCTGCCGATGCTGGTTATCAGCCCGTATGCAAAAGCTAACTACGTTGACCACTCTCTGCTGAATCAGGCGTCTGTCCTGAAATTCATCGAATACAACTGGGGTATCGGTTCCGTATCTAAATACTCTAACGACAAGTATTCTAACAATATCCTGAATATGTTTGACTTCAACAAAGAGCAAAAAACCCTGAAACTGATCCTGGACCCGAAAACTGGTCTCGTTATGGATAAACTGAACTAA *GC% Content for gene: 47.77%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
TACTTCCAATCCATGGACGTGAATAATTCTAAGCCGAACGATTACGGTACCCTGGTTAAAATCGAACAGAAACT
GTTTAACAACGCGAATACGCTCAAAACGACCACCCCGATTAAACACGTAGTCATTATCTTCCAGGAGAATAACT
CTTTTGATCGTTACTTCGGTATGTACCCGAATGCGAAGAATCCGGAAGGTGAACCAAAGTTTGTTGCGAAAGA
AAACACCCCAAATGTTAACGGCCTCACCAAACAGCTGCTCGAAAATAACCCGAACACCAAAAATCCGTACCGC
CTGGATCGCAACTTCCAGCCGTGCTCTCAGAACCACGAATACCACCAGGAAATCTCTTCTTTCAATGGTGGCCT
GATGAATAAGTTCGTCGAACATGGTGGTCACGACAATGACACCTACAAACAGAATTGCGATGGTCAGGTGAT
GGGCTACTATGATGGCAATACTGTTACCGCGCTGTGGAATTACGCGCAGAACTTCGCGCTGAATGACAACAC
CTTTGGTACTACCTTCGGTCCGTCTACCCCGGGTGCCCTGAACCTGGTTGCGGGTGCAAACGGTCCGGCGAT
GTCTCCGTCTGGCAACCTCGAGAACATCGAAAACAACTACATCATTGACGACCCTAACCCGTATTATGACGACT
GCTCTTACGGCACCTCTAAATCTGGCGACACCAACACTGCGGTTGCTAAGATCACGGATGGTTACAACATCGG
TCACTACCTCACCCAAAAAGGTATCACTTGGGGTTGGTTTCAAGGCGGCTTCAAACCGACCTCTTACTCTGGTA
AGACGGCGATTTGTGACGCCATGTCTACCAACAAGTTCGGTGTTAAATCTCGTGACTATATCCCGCACCACGA
GCCTTTCAACTATTGGAAGGAGACCTCCAATCCACACCACCTCGCGCCGAGCGACGACAAATACATTGGTTCT
AATGATCAAGCGAATCACCAATACGACATCTCTGAATTCTGGAAGGCGCTCGACCAGAACAACATGCCTGCCG
TTTCTTACCTGAAAGCTCCGGGTTACCAGGATGGCCACGGTGGCTACTCCAACCCACTGGACGAACAGGAAT
GGCTCGTTAATACCATCAACCGTATCCAGCAGTCTAAAGACTGGGACTCTACCGCTATTATCATCATTTACGAT
GACAGCGACGGTGATTACGATCACGTTTACTCTCCAAAGTCTCAATTCTCTGACATCAAGGGCCGTCAAGGTT
ATGGTCCGCGTCTGCCGATGCTGGTTATCAGCCCGTATGCAAAAGCTAACTACGTTGACCACTCTCTGCTGAA
TCAGGCGTCTGTCCTGAAATTCATCGAATACAACTGGGGTATCGGTTCCGTATCTAAATACTCTAACGACAAGT
ATTCTAACAATATCCTGAATATGTTTGACTTCAACAAAGAGCAAAAAACCCTGAAACTGATCCTGGACCCGAAA
ACTGGTCTCGTTATGGATAAACTGAACTAACAGTAAAGGTGGATA
*GC% Content for gene (codon optimized): 47.65%
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): 38 total
*NCBI Gene # or RefSeq#: FTN_0090
*Protein ID (NP or XP #) or Wolbachia#: YP_897755.1
*Organism (including strain): Francisella tularensis
Etiologic Risk Group: Those who live or visit areas of Arkansas, Missouri, or Oklahoma due to the concentration of ticks. Also those who engage in hunting, trapping, gardening, landscaping, or work in wildlife management or veterinary medicine.
*Disease Information (sort of like the Intro to your Mini Research Write up):
Tularemia is an infectious disease caused by the bacteria Francisella tularensis, commonly found in animals such as rodents, rabbits and hares [4]. Humans can contract tularemia by means of getting bitten by an infected tick, deerfly, or other insect, handling an infected animal carcass, eating or drinking contaminated food or water, or breathing in the bacteria itself [4]. Tularemia ranges from mild to life-threatening, often with the accompanying symptoms of a sudden fever, chills, headaches, diarrhea, muscle aches, joint paint, dry cough, and progressive weakness [4]. More serious symptoms include development of pneumonia, chest pain, cough and difficulty breathing [4]. Tularemia occurs naturally in the United States and because of its highly infectious nature, it can be used as a bioterrorism weapon.
Link to TDR Targets page (if present): n/a
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)
http://www.ncbi.nlm.nih.gov/gene/?term=FTN_0090
Essentiality of this protein: Essential in producing, acquiring, and mobilizing inorganic phosphates in the cell signaling transduction pathway
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): 2-mer
Complex of proteins?: complexed with othrovanadate (2D1G)
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): No evidence showing specific druggability of AcpA in another organism, but there are papers showing druggability of secreted phosphatases in other organisms
http://www.researchgate.net/profile/Yossef_Av-Gay/publication/232529387_Mycobacterium_tuberculosis-secreted_phosphatases_from_pathogenesis_to_targets_for_TB_drug_development/links/0deec5203d3ebdc8f6000000.pdf
*EC#: 3.1.3.2
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/enzyme.php?ecno=3.1.3.2&Suchword=&organism%5B%5D=Francisella+tularensis
-- Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
A discontinuous colorimetric assay was used. Standard 0.2 ml assay mixture contained 0.2 M sodium acetate, pH 6.0, 2.0 mM p-nitrophenylphosphate (pNPP), and varying amounts of enzyme. Mixtures incubated at 37oC for 15 minutes, and reaction was stopped by addition of 0.2 ml of 0.5 M glycine, pH 10.
-- link to Sigma (or other company) page for assay (see Sigma links below)
-- -or link (or citation) to paper that contains assay information
http://www.sciencedirect.com/science/article/pii/S1046592805001580
-- links to assay reagents (substrates) pages.
http://www.sigmaaldrich.com/catalog/product/sigma/g7126?lang=en®ion=US
--- List cost and quantity of substrate reagents, supplier, and catalog #
Glycine
Several different sizes available, see site above for complete list.
Supplier:ReagentPlus®
10 mg = $17.80
100 g = $23.50
1 kg = $64.50
Structure (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 2D1G
Current Inhibitors:
AgNO3, Arsenate, Cu2+, FeSO4, Hg2+, Molybdate, Orthovanadate, Tartrate, Tungstate, Vanadate, Zn2+
Expression Information (has it been expressed in bacterial cells):
Expressed in E. coli BL21 (DE3)
Purification Method:
SP-Sepharose cation exchange resin (25 ml)
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): (NOT INCLUDING ADDED X)
MKLNKITLGILSLSIATTTFATDVNNSKPNDYGTLVKIEQKLFNNANTLKTTTPIKHVVIIFQENNSFDRYFGMYPN
AKNPEGEPKFVAKENTPNVNGLTKQLLENNPNTKNPYRLDRNFQPCSQNHEYHQEISSFNGGLMNKFVEHG
GHDNDTYKQNCDGQVMGYYDGNTVTALWNYAQNFALNDNTFGTTFGPSTPGALNLVAGANGPAMSPSGNL
ENIENNYIIDDPNPYYDDCSYGTSKSGDTNTAVAKITDGYNIGHYLTQKGITWGWFQGGFKPTSYSGKTAICDA
MSTNKFGVKSRDYIPHHEPFNYWKETSNPHHLAPSDDKYIGSNDQANHQYDISEFWKALDQNNMPAVSYLKA
PGYQDGHGGYSNPLDEQEWLVNTINRIQQSKDWDSTAIIIIYDDSDGDYDHVYSPKSQFSDIKGRQGYGPRLP
MLVISPYAKANYVDHSLLNQASVLKFIEYNWGIGSVSKYSNDKYSNNILNMFDFNKEQKTLKLILDPKTGLVMD
KLN
*length of your protein in Amino Acids: 514
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website:
57.78 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 91930
Abs 0.1% (=1 g/l) 1.591, assuming all pairs of Cys residues form cystines
Ext. coefficient 91680
Abs 0.1% (=1 g/l) 1.587, assuming all Cys residues are reduced
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
This present two possible trans-membrane regions and one definite region. Therefore there is 1-3 trans-membrane regions.
*CDS Gene Sequence (paste as text only):
ATGGACGTGAATAATTCTAAGCCGAACGATTACGGTACCCTGGTTAAAATCGAACAGAAACTGTTTAACAACGCGAATACGCTCAAAACGACCACCCCGATTAAACACGTAGTCATTATCTTCCAGGAGAATAACTCTTTTGATCGTTACTTCGGTATGTACCCGAATGCGAAGAATCCGGAAGGTGAACCAAAGTTTGTTGCGAAAGAAAACACCCCAAATGTTAACGGCCTCACCAAACAGCTGCTCGAAAATAACCCGAACACCAAAAATCCGTACCGCCTGGATCGCAACTTCCAGCCGTGCTCTCAGAACCACGAATACCACCAGGAAATCTCTTCTTTCAATGGTGGCCTGATGAATAAGTTCGTCGAACATGGTGGTCACGACAATGACACCTACAAACAGAATTGCGATGGTCAGGTGATGGGCTACTATGATGGCAATACTGTTACCGCGCTGTGGAATTACGCGCAGAACTTCGCGCTGAATGACAACACCTTTGGTACTACCTTCGGTCCGTCTACCCCGGGTGCCCTGAACCTGGTTGCGGGTGCAAACGGTCCGGCGATGTCTCCGTCTGGCAACCTCGAGAACATCGAAAACAACTACATCATTGACGACCCTAACCCGTATTATGACGACTGCTCTTACGGCACCTCTAAATCTGGCGACACCAACACTGCGGTTGCTAAGATCACGGATGGTTACAACATCGGTCACTACCTCACCCAAAAAGGTATCACTTGGGGTTGGTTTCAAGGCGGCTTCAAACCGACCTCTTACTCTGGTAAGACGGCGATTTGTGACGCCATGTCTACCAACAAGTTCGGTGTTAAATCTCGTGACTATATCCCGCACCACGAGCCTTTCAACTATTGGAAGGAGACCTCCAATCCACACCACCTCGCGCCGAGCGACGACAAATACATTGGTTCTAATGATCAAGCGAATCACCAATACGACATCTCTGAATTCTGGAAGGCGCTCGACCAGAACAACATGCCTGCCGTTTCTTACCTGAAAGCTCCGGGTTACCAGGATGGCCACGGTGGCTACTCCAACCCACTGGACGAACAGGAATGGCTCGTTAATACCATCAACCGTATCCAGCAGTCTAAAGACTGGGACTCTACCGCTATTATCATCATTTACGATGACAGCGACGGTGATTACGATCACGTTTACTCTCCAAAGTCTCAATTCTCTGACATCAAGGGCCGTCAAGGTTATGGTCCGCGTCTGCCGATGCTGGTTATCAGCCCGTATGCAAAAGCTAACTACGTTGACCACTCTCTGCTGAATCAGGCGTCTGTCCTGAAATTCATCGAATACAACTGGGGTATCGGTTCCGTATCTAAATACTCTAACGACAAGTATTCTAACAATATCCTGAATATGTTTGACTTCAACAAAGAGCAAAAAACCCTGAAACTGATCCTGGACCCGAAAACTGGTCTCGTTATGGATAAACTGAACTAA
*GC% Content for gene: 47.77%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
TACTTCCAATCCATGGACGTGAATAATTCTAAGCCGAACGATTACGGTACCCTGGTTAAAATCGAACAGAAACT
GTTTAACAACGCGAATACGCTCAAAACGACCACCCCGATTAAACACGTAGTCATTATCTTCCAGGAGAATAACT
CTTTTGATCGTTACTTCGGTATGTACCCGAATGCGAAGAATCCGGAAGGTGAACCAAAGTTTGTTGCGAAAGA
AAACACCCCAAATGTTAACGGCCTCACCAAACAGCTGCTCGAAAATAACCCGAACACCAAAAATCCGTACCGC
CTGGATCGCAACTTCCAGCCGTGCTCTCAGAACCACGAATACCACCAGGAAATCTCTTCTTTCAATGGTGGCCT
GATGAATAAGTTCGTCGAACATGGTGGTCACGACAATGACACCTACAAACAGAATTGCGATGGTCAGGTGAT
GGGCTACTATGATGGCAATACTGTTACCGCGCTGTGGAATTACGCGCAGAACTTCGCGCTGAATGACAACAC
CTTTGGTACTACCTTCGGTCCGTCTACCCCGGGTGCCCTGAACCTGGTTGCGGGTGCAAACGGTCCGGCGAT
GTCTCCGTCTGGCAACCTCGAGAACATCGAAAACAACTACATCATTGACGACCCTAACCCGTATTATGACGACT
GCTCTTACGGCACCTCTAAATCTGGCGACACCAACACTGCGGTTGCTAAGATCACGGATGGTTACAACATCGG
TCACTACCTCACCCAAAAAGGTATCACTTGGGGTTGGTTTCAAGGCGGCTTCAAACCGACCTCTTACTCTGGTA
AGACGGCGATTTGTGACGCCATGTCTACCAACAAGTTCGGTGTTAAATCTCGTGACTATATCCCGCACCACGA
GCCTTTCAACTATTGGAAGGAGACCTCCAATCCACACCACCTCGCGCCGAGCGACGACAAATACATTGGTTCT
AATGATCAAGCGAATCACCAATACGACATCTCTGAATTCTGGAAGGCGCTCGACCAGAACAACATGCCTGCCG
TTTCTTACCTGAAAGCTCCGGGTTACCAGGATGGCCACGGTGGCTACTCCAACCCACTGGACGAACAGGAAT
GGCTCGTTAATACCATCAACCGTATCCAGCAGTCTAAAGACTGGGACTCTACCGCTATTATCATCATTTACGAT
GACAGCGACGGTGATTACGATCACGTTTACTCTCCAAAGTCTCAATTCTCTGACATCAAGGGCCGTCAAGGTT
ATGGTCCGCGTCTGCCGATGCTGGTTATCAGCCCGTATGCAAAAGCTAACTACGTTGACCACTCTCTGCTGAA
TCAGGCGTCTGTCCTGAAATTCATCGAATACAACTGGGGTATCGGTTCCGTATCTAAATACTCTAACGACAAGT
ATTCTAACAATATCCTGAATATGTTTGACTTCAACAAAGAGCAAAAAACCCTGAAACTGATCCTGGACCCGAAA
ACTGGTCTCGTTATGGATAAACTGAACTAACAGTAAAGGTGGATA
*GC% Content for gene (codon optimized): 47.65%
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): 38 total
DNA Works Output:
File Link
Primer design results for 'tail' primers (this is just 2 sequences):
Forward Primer:
5’ TAC TTC CAA TCC ATGGAC GTG AAT AAT TCT 3’ 30 bp
GC Content 36.7%
Reverse Primer:
5’ TAT CCA CCT TTA CTG TTA GTT CAG TTT ATC CAT 3’
GC Content 33.3%